Expression Of Immune Checkpoint Markers Research Articles

Assessment of Immune Status in Patients with Mismatch Repair Deficiency Endometrial Cancer.

This study introduced a novel subtype classification method for endometrial cancer (EC) with mismatch repair deficiency (MMRd) by employing immune status and prognosis as the foundational criteria. The goal was to enhance treatment guidance through precise subtype delineation. Study Cohort: This study encompassed a cohort of 119 patients diagnosed with MMRd-EC between 2015 and 2022. Analyses using t-tests and Mann-Whitney U-tests were performed to assess prognostic markers and peripheral blood immune cell profiles in patients with MutS deficiency (MutS-d) versus those with MutL deficiency (MutL-d). Logistic regression analysis was used to identify independent risk factors. Bioinformatics Analysis: An online database was used to assess the prognostic implications, immune cell infiltration, and immune checkpoint involvement associated with the deficiency of MutS versus MutL in EC. Patients with MutL-d exhibited heightened risk factors, including elevated cancer grade and increased myometrial invasion, leading to poorer prognosis and shorter overall survival and progression-free survival. Regarding systemic immune status, patients with MutL-d demonstrated decreased peripheral blood lymphocyte percentage, lymphocyte count, and CD8+ T cell percentage. For local immunity, the infiltration of natural killer cells, CD8+ T cells, and cytotoxic T lymphocytes in the tumor tissue was reduced in patients with MutL-d. Additionally, patients with MutL-d exhibited lower expression of immune checkpoint markers. The composition of immune subtypes and survival outcomes also indicate that patients with MutL-d have a poorer immune status and prognosis than the patients with MutS-d. Patients with MMRd-EC can be subclassified according to MutS or MutL deficiency. Patients with MutS-d exhibited better immune status, prognosis, and immunotherapy benefits than those with MutL-d. These results can help guide patients to a more precise treatment.

KEAP1-Mutant Lung Cancers Weaken Anti-Tumor Immunity and Promote an M2-like Macrophage Phenotype.

Considerable advances have been made in lung cancer therapies, but there is still an unmet clinical need to improve survival for lung cancer patients. Immunotherapies have improved survival, although only 20-30% of patients respond to these treatments. Interestingly, cancers with mutations in Kelch-like ECH-associated protein 1 (KEAP1), the negative regulator of the nuclear factor erythroid 2-related factor 2 (NRF2) transcription factor, are resistant to immune checkpoint inhibition and correlate with decreased lymphoid cell infiltration. NRF2 is known for promoting an anti-inflammatory phenotype when activated in immune cells, but the study of NRF2 activation in cancer cells has not been adequately assessed. The objective of this study was to determine how lung cancer cells with constitutive NRF2 activity interact with the immune microenvironment to promote cancer progression. To assess, we generated CRISPR-edited mouse lung cancer cell lines by knocking out the KEAP1 or NFE2L2 genes and utilized a publicly available single-cell dataset through the Gene Expression Omnibus to investigate tumor/immune cell interactions. We show here that KEAP1-mutant cancers promote immunosuppression of the tumor microenvironment. Our data suggest KEAP1 deletion is sufficient to alter the secretion of cytokines, increase expression of immune checkpoint markers on cancer cells, and alter recruitment and differential polarization of immunosuppressive macrophages that ultimately lead to T-cell suppression.

Deciphering m6A signatures in hepatocellular carcinoma: Single-cell insights, immune landscape, and the protective role of IGFBP3.

RNA m6 methyladenosine (m6A) modifications impact tumor biology and immune processes, particularly in hepatocellular malignant tumors. Using a consensus clustering algorithm on 371 hepatocellular carcinoma (HCC) samples, we identified three m6A-modified subtypes and correlated them with positive tumor microenvironment (TME) markers for distinct immune phenotypes. Stratifying patients based on m6A scores revealed a low presentation group with better immune penetration, lower tumor mutation load, and increased expression of immune checkpoint markers like CTLA-4 and PD-1, suggesting enhanced responsiveness to immunization therapy. A machine-learning model of 23 m6A genes was constructed. Single-cell analysis revealed a surprising enrichment of IGFBP3 in astrocytes, prompting the exploration of associated signaling pathways. Experimental verification shows that IGFBP3 is significantly enhanced in normal tissues, while immunohistochemical analysis shows that its expression is lower in tumor tissues, indicating its protective effect in HCC and a good prognosis. Importantly, high IGFBP3 expression is associated with better outcomes in patients receiving immunotherapy. Moreover, cytotoxic T lymphocyte (CTL) experiments have confirmed that high expression of IGFBP3 is associated with stronger T cell-killing ability. In summary, the comprehensive evaluation of m6A modification, immune characteristics, and single-cell analysis in this study not only revealed the TME of HCC but also made significant contributions to the progress of personalized HCC immunotherapy targeting IGFBP3. This study provides a solid theoretical foundation for clinical translation and emphasizes its potential impact on developing effective treatment strategies.

γδ T cells mediate robust anti-HIV functions during antiretroviral therapy regardless of immune checkpoint expression.

Although antiretroviral therapy (ART) efficiently suppresses HIV viral load, immune dysregulation and dysfunction persist in people living with HIV (PLWH). γδ T cells are functionally impaired during untreated HIV infection, but the extent to which they are reconstituted upon ART is currently unclear. Utilising a cohort of ART-treated PLWH, we assessed the frequency and phenotype, characterised in vitro functional responses and defined the impact of immune checkpoint marker expression on effector functions of both Vδ1 and Vδ2 T cells. We additionally explore the in vitro expansion of Vδ2 T cells from PLWH on ART and the mechanisms by which such expanded cells may sense and kill HIV-infected targets. A matured NK cell-like phenotype was observed for Vδ1 T cells among 25 ART-treated individuals (PLWH/ART) studied compared to 17 HIV-uninfected controls, with heightened expression of 2B4, CD160, TIGIT and Tim-3. Despite persistent phenotypic perturbations, Vδ1 T cells from PLWH/ART exhibited strong CD16-mediated activation and degranulation, which were suppressed upon Tim-3 and TIGIT crosslinking. Vδ2 T cell degranulation responses to the phosphoantigen (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate at concentrations up to 2 ng mL-1 were significantly impaired in an immune checkpoint-independent manner among ART-treated participants. Nonetheless, expanded Vδ2 T cells from PLWH/ART retained potent anti-HIV effector functions, with the NKG2D receptor contributing substantially to the elimination of infected cells. Our findings highlight that although significant perturbations remain within the γδ T cell compartment throughout ART-treated HIV, both subsets retain the capacity for robust anti-HIV effector functions.

IMMU-25. ANALYSIS OF IMMUNOSUPPRESSIVE MYELOID CELLS IN THE GLIOBLASTOMA TUMOR MICROENVIRONMENT REVEALS THERAPEUTICALLY TARGETABLE IMMUNOMODULATORY SIGNALS ACROSS POPULATIONS

Abstract Immunosuppression in glioblastoma (GBM) remains a critical and poorly understood barrier to the development of effective novel therapies. Increasing evidence points to the critical role of immunosuppressive myeloid cells in the immunosuppressive tumor microenvironment and an intriguing target for ameliorating tumor-mediated immune suppression. Efforts to characterize the myeloid milieu in the tumor microenvironment have revealed a highly heterogenous population of cells with the capacity to exert immunosuppressive effects. In order to maximize the efficacy of potential therapeutic targets, we sought to identify immune checkpoint markers expressed across a range of immunomodulatory myeloid cell populations in the tumor microenvironment of patients undergoing resection for GBM. Freshly collected ultrasonic aspirate obtained during the primary surgery for GBM resection (based on preoperative imaging and intraoperative frozen section, later confirmed on final pathology) was prepared for high dimensional flow cytometry to quantify the immune checkpoint marker expression profile of a number of described immunosuppressive myeloid cell populations, including both monocytic and granulocytic myeloid-derived suppressor cells (mMDSC and gMDSC), and CD16+ non-classical monocytes (NCM). mMDSCs expressed particularly high levels of TIGIT (53.9±19.27%), which was also highly expressed in the NCM population (25.03±5.44%). NCMs also demonstrated high levels of B7H3 expression (39.85±18.05%), which has recently been implicated as an important immune checkpoint protein in GBM. gMDSCs demonstrated a rather different expression profile, with significantly higher levels of VISTA expression than other immunomodulatory markers (p< 0.05). These findings underscore the diversity of immunomodulatory signaling in the tumor-associated myeloid cell compartment, emphasizing the challenges in developing single-target immune checkpoint inhibitor therapy in GBM. Nevertheless, this data points to some common patterns of expression across major immunomodulatory cell populations that remain understudied in GBM and could herald more promising avenues of immunotherapy development.

Deciphering Treg cell roles in esophageal squamous cell carcinoma: a comprehensive prognostic and immunotherapeutic analysis.

Background: Esophageal squamous cell carcinoma (ESCC) is a prevalent and aggressive form of cancer that poses significant challenges in terms of prognosis and treatment. Regulatory T cells (Treg cells) have gained attention due to their influential role in immune modulation within the tumor microenvironment (TME). Understanding the intricate interactions between Treg cells and the tumor microenvironment is essential for unraveling the mechanisms underlying ESCC progression and for developing effective prognostic models and immunotherapeutic strategies. Methods: A combination of single-cell RNA sequencing (scRNA-seq) and bulk RNA-seq analysis was utilized to explore the role of Treg cells within the TME of ESCC. The accuracy and applicability of the prognostic model were assessed through multi-dimensional evaluations, encompassing an examination of the model's performance across various dimensions, such as the mutation landscape, clinical relevance, enrichment analysis, and potential implications for immunotherapy strategies. Results: The pivotal role of the macrophage migration inhibitory factor (MIF) signaling pathway within the ESCC TME was investigated, with a focus on its impact on Treg cells and other subpopulations. Through comprehensive integration of bulk sequencing data, a Treg-associated signature (TAS) was constructed, revealing that ESCC patients with elevated TAS (referred to as high-TAS individuals) experienced significantly improved prognoses. Heightened immune infiltration and increased expression of immune checkpoint markers were observed in high-TAS specimens. The model's validity was established through the IMvigor210 dataset, demonstrating its robustness in predicting prognosis and responsiveness to immunotherapy. Heightened therapeutic benefits were observed in immune-based interventions for high-TAS ESCC patients. Noteworthy differences in pathway enrichment patterns emerged between high and low-TAS cohorts, highlighting potential avenues for therapeutic exploration. Furthermore, the clinical relevance of key model genes was substantiated by analyzing clinical samples from ten paired tumor and adjacent tissues, revealing differential expression levels. Conclusion: The study established a TAS that enables accurate prediction of patient prognosis and responsiveness to immunotherapy. This achievement holds significant implications for the clinical management of ESCC, offering valuable insights for informed therapeutic interventions.

Comparison of immune microenvironment between primary and metastatic breast tumors.

1030 Background: Immune evasion is one of the mechanisms by which cancer cells gain the ability to metastasize from the primary tumor to distant sites. In triple-negative breast cancer (TNBC), metastatic tumors are shown to be more immunologically silent than primary tumors. As a result, there are varying degrees of responses to immunotherapy between early-stage and metastatic tumors. Positive clinical responses to immune checkpoint inhibitors (ICIs) are seen in patients with early-stage TNBC regardless of PD-L1 expression. In this study, we investigated the differences in the immune signatures of primary and metastatic breast cancer in a real-world patient population. Methods: We analyzed retrospective comprehensive genomic and immune profiling (CGIP) results from 529 breast tumors tested in the real-world clinical setting. Tumor specimens were classified as primary breast (PB), any lymph nodes (LN; regional and non-regional) or metastatic visceral (MV) sites. LN samples were chosen as positive controls due to expected elevated inflammatory signaling. PD-L1 (CPS positive ≥10) and TMB (high ≥10 Mut/Mb) were determined using IHC and DNA sequencing, respectively. mRNA expression signatures of tumor inflammation (TIGS, strong/moderate/weak) and expression of LAG3, TIGIT and TIM3 were determined by RNA-sequencing from a 395-gene panel. LAG3, TIGIT and TIM3 were selected due to emerging clinical trials in solid tumors. We used over-representation and proportion analysis using chi-squared test to determine the association of specimen sites to various genomic and immune correlates. Results: Among 529 cases, the median patient age was 63.2 years (25.5-93.5). The majority of patients were female (519, 98%), and the most common tumor histology was invasive ductal carcinoma (287, 54%). A total of 224 (42%), 72 (14%), and 232 (44%) of patients had specimens from PB, LN, MV, respectively, while 1 (0.19%) patient had an unknown source. Samples of PB harbored greater degree of immune infiltration, demonstrating higher TIGS score than MV (p=0.014). Primary lesions also demonstrated a greater proportion of PD-L1 positive than metastatic lesions (44% vs 21%, p < 0.001) and higher expressions of immune checkpoints such as TIGIT (p<0.001), LAG3 (p=0.037) and TIM3 (p<0.001). Conclusions: We found that non-lymph node breast cancer metastases harbor a less active immune response than primary breast lesions, showing a lower degree of immune cell infiltration and decreased expression of immune checkpoint markers. These findings support the notion that the immune microenvironment of breast cancer metastases is immunosuppressive and may exhibit a tempered response to ICIs. Therefore, combination treatments of ICIs with chemotherapy, targeted therapies or cancer vaccines may be promising therapeutic approaches to enhance the immune responses and potentially overcome resistance to ICIs in metastatic breast cancer.

Comparative expression analysis of immune-related markers in surgically resected lung neuroendocrine neoplasms

BackgroundAlthough immunotherapy has led to a paradigm shift in the treatment of lung cancer, the therapeutic approaches for lung neuroendocrine neoplasms (LNENs) are still limited. Our aim was to explore the immunological landscape and the expression of immune checkpoint markers in LNENs. MethodsSurgically removed tumor samples of 26 atypical carcinoid (AC), 30 large cell neuroendocrine carcinoma (LCNEC) and 29 small cell lung cancer (SCLC) patients were included. The immune phenotype of each tumor type was assessed by using a panel of 15 immune-related markers. As these markers are potentially expressed by immune cells and/or tumor cells, they might serve as putative targets for immunotherapy. Expression patterns were measured by immunohistochemistry and correlated with clinicopathological parameters and prognosis. ResultsUnsupervised hierarchical clustering revealed distinct immunologic profiles across tumor types. Specifically, AC tumors were characterized by high tumor cell CD40 expression and low levels of immune infiltrates whereas SCLC samples had a high CD47 and Inducible T Cell Costimulator (ICOS) expression in tumor cells and immune cells, respectively. High CD70 and CD137 expression by tumor cells as well as elevated expression of CD27, Lymphocyte Activation Gene 3 (LAG3), and CD40 by immune cells were characteristic for LCNEC samples. Overall, SCLC and LCNEC tumors had a more immunogenic phenotype than AC samples. High tumor cell CD47 and CD40 expressions were associated with impaired and improved survival outcomes, respectively. ConclusionsBy providing insights into the widely divergent immunologic profiles of LNENs, our results might serve as a basis for the development of novel immunotherapy-related approaches in these devastating malignancies.

Abstract 3268: Biomarkers of drug-tolerant persistent melanoma cells in human, correlation with tumor microenvironment and response to treatment

Abstract Introduction: Drug tolerant cancer persister cells can be silent for years but constitute a reservoir of genetic resistance and can eventually result in tumor relapse. We showed that targeted therapy (TT)-tolerant BRAFV600 melanoma persister cells (MPC) have a reduced global mRNA translation activity but an increased translation efficiency of a subset of mRNAs encoding proteins involved in transcription, chromatin remodeling and epigenetic changes. Here, we identify protein markers of MPC in patient biopsies treated either with TT or with immune checkpoint inhibitors (ICI). Since persister cells are not destroyed by the immune system, we explored the interactions of MPC with their immune microenvironment. Single cell expression of these candidate MPC markers, as well markers of the tumor microenvironment (31 lymphoid and myeloid markers), were studied in sequential biopsies from melanoma patients treated with TT or ICI by CyTOF. Spatial localization of MPC markers and various types of immune cells were studied by multiplex immunohistochemistry (Akoya Codex). In addition, the in vitro effect of the cell supernatants of MPC or naïve parental cells on immune cells was studied. Results: Biomarkers of persistence included 7 protein markers that are regulated at the translational level (MLL3, HDAC6, CCSER2, MT-ND5, ANK2, HIPK2, CBP) and 4 that are transcriptionally regulated (MITF, NGFR, CD36 and SLIT2). Some MPC markers were expressed at baseline in melanoma patients’ biopsies (n = 27) and their baseline expression tended to be associated with progressive disease (to TT or ICI). Paired biopsies from 18 patients showed that an increase in HDAC6, ANK2, HIPK2, CBP5 and MITF was statistically correlated with a short survival regardless of the type of treatment. Baseline lymphocytic infiltrate, especially CD4+ T cells, was associated with favorable clinical outcome. In addition, we found that the supernatants derived from naïve parental and persister cells have distinct effects on human CD4+, CD8+ T cells and macrophages. MPC supernatant had a significant inhibitory effect on the proliferation and activation of CD4+, CD8+ T cells as well as the expression of immune checkpoint markers (PD-1, PD-L1, CTLA4, LAG3, Tim3) compared to supernatant of naïve parental cells. We also found that supernatants of MPC induced M2-like polarization of macrophages. One major candidate to convey this effect is MIF (Macrophage migration inhibitory factor) that is specifically increased in MPC supernatant. Conclusion: Melanoma persister cells can be identified in patients’ biopsies during TT as well as ICI therapy. The expression of MPC protein markers is associated with poor clinical outcome regardless of the type of treatment. Their expression is inversely correlated with a hot immune environment that could result from the effect of immunosuppressive cytokines secreted by persister cells. Citation Format: Hongxu Chen, Shensi Shen, Sara Faouzi, Alexia Alfaro, Dorothee Baille, Séverine Roy, Naima Benannoune, Hugues Hermann, Samad Muhammadnejad, Stephan Vagner, Caroline Robert. Biomarkers of drug-tolerant persistent melanoma cells in human, correlation with tumor microenvironment and response to treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3268.

Tumor-Infiltrating T Cells in EBV-Associated Gastric Carcinomas Exhibit High Levels of Multiple Markers of Activation, Effector Gene Expression, and Exhaustion

Epstein-Barr virus (EBV) is a gamma-herpesvirus associated with 10% of all gastric cancers (GCs) and 1.5% of all human cancers. EBV-associated GCs (EBVaGCs) are pathologically and clinically distinct entities from EBV-negative GCs (EBVnGCs), with EBVaGCs exhibiting differential molecular pathology, treatment response, and patient prognosis. However, the tumor immune landscape of EBVaGC has not been well explored. In this study, a systemic and comprehensive analysis of gene expression and immune landscape features was performed for both EBVaGC and EBVnGC. EBVaGCs exhibited many aspects of a T cell-inflamed phenotype, with greater T and NK cell infiltration, increased expression of immune checkpoint markers (BTLA, CD96, CTLA4, LAG3, PD1, TIGIT, and TIM3), and multiple T cell effector molecules in comparison with EBVnGCs. EBVaGCs also displayed a higher expression of anti-tumor immunity factors (PDL1, CD155, CEACAM1, galectin-9, and IDO1). Six EBV-encoded miRNAs (miR-BARTs 8-3p, 9-5p, 10-3p, 22, 5-5p, and 14-3p) were strongly negatively correlated with the expression of immune checkpoint receptors and multiple markers of anti-tumor immunity. These profound differences in the tumor immune landscape between EBVaGCs and EBVnGCs may help explain some of the observed differences in pathological and clinical outcomes, with an EBV-positive status possibly being a potential biomarker for the application of immunotherapy in GC.

765P Comprehensive analysis of the association between RAS mutation and immune checkpoint marker expression

The effectiveness of KRAS G12C inhibition as a cancer treatment was recently reported. Knowing the association between RAS mutation and the expression level of immune checkpoint markers is of interest since it may guide a potential combination therapy of RAS inhibition and immune checkpoint blockade.

Comprehensive transcriptomic analysis of immune checkpoint markers in a pancancer cohort: Implications for response and resistance.

2555 Background: Although immune checkpoint blockade (ICB) has revolutionized cancer treatment, not all patients with cancer benefit from ICB. One possible explanation for poor responders/resistance is the variable expression level of the target molecules (e.g., PD-1 and PD-L1) in the tumor microenvironment. There are recent or ongoing trials targeting variable pathways for immune evasion (e.g., LAG3 or IDO1). It is therefore of interest to know the expression levels related to variable immune checkpoints so that clinical trials can focus on the patients who can benefit from the cognate treatment. Methods: Overall, 514 patients with various solid tumors seen at the University of San Diego, Moores Center for Personalized Cancer Therapy were analyzed. The expression levels of checkpoint markers (ADORA2A, BTLA, CD276, CTLA4, IDO1, IDO2, LAG3, NOS2, PD-1, PD-L1, PD-L2, PVR, TIGIT, TIM3, VISTA, and VTCN) in the tumor samples were measured through RNA sequencing and normalized to internal housekeeping gene profiles, and ranked from 0 to 100 percentile based on a reference population. The expressions of each checkpoint marker were correlated with cancer types, microsatellite instability (MSI), tumor mutational burden (TMB), and programmed death-ligand 1 (PD-L1) status on immunohistochemistry. Results: In this cohort, 60% were female, median age of 60, and included 30 different tumor types, with colorectal cancer being the most common (27%). The rank values of all checkpoint markers were distributed broadly from 0 to 99 or 100. CD276 and NOS2 had the highest (68th percentile) and lowest (13.5 percentile) median rank values, respectively. When rank values were categorized to “Low” (0-24), “Intermediate” (25-74), and “High” (75-100), 41.6% of patients showed high expression of CD276 while only 13% showed high expression of PD-L1. Each patient had a distinctive protfolio of the categorical expression levels of 16 checkpoint markers. Several checkpoint markers, especially NOS2, showed a significant correlation with cancer type. (median rank values in colorectal, stomach, pancreatic, and breast cancer were 79, 76, 5 and 0 respectively, p < 0.001) Five markers (IDO1, LAG3, PD-1, PD-L1, and TIGIT) showed significant correlation with MSI, while seven markers (CTLA4, IDO1, LAG3, PD-1, PD-L1, PD-L2, and TIGIT) were significantly associated with positive PD-L1 status. However, no significant association was seen based on TMB or tissue-specific grouping of patients. Conclusions: The expression of immune checkpoint markers varies from patient to patient, though transcript expression of several markers correlates with cancer type, MSI, and PD-L1 status. Clinical trials with patient selection based on the expression level of checkpoint markers matched to the corresponding ICB drug are warranted.

Pan-cancer analysis reveals RIPK2 predicts prognosis and promotes immune therapy resistance via triggering cytotoxic T lymphocytes dysfunction

BackgroundReceptor-interacting protein kinase 2 (RIPK2, also known as RIP2) was reported to be associated with bacterial infections as well as inflammatory responses. However, the role of RIPK2 in prognosis and immunotherapy response is yet to be elucidated in human pan-cancer.MethodsIn this study, we investigated the expression, gene alteration landscape and prognostic value of RIPK2 in 33 cancers through various databases including Ualcan, cBioportal and Gene Expression Profiling Interactive Analysis 2 (GEPIA2). Then, the correlation between RIPK2 and immune infiltration, immune score, stromal score, and ESTIMATE score was investigated in the Cancer Genome Atlas (TCGA) and tumor immune estimation resource (TIMER) databases. Independent cohorts were utilized to explore the role of RIPK2 in tumor immunotherapy response. Furthermore, Gene set enrichment analysis (GSEA) was conducted to explore the mechanisms by which RIPK2 regulates immune therapy resistance. Single-cell RNA-seq datasets were used to analyze the expression level of RIPK2 on different immune cells. Moreover, CellMiner database was used to explore the relationship between RIPK2 expression with drug response.ResultCompared with normal tissue, tumor tissue had a higher expression level of RIPK2 in various cancers. Survival analysis showed that high expression of RIPK2 associated with poor prognosis in numerous cancers. RIPK2 was found to promote a series of immune cell infiltration and B cells, macrophages, and neutrophils were significantly positively correlated with the expression of RIPK2. Moreover, RIPK2 affected immune score, stromal score and ESTIMATE score for a wide range of cancers. In the vast majority of 33 cancers, gene co-expression analysis showed that RIPK2 was positively correlated with the expression of immune checkpoint markers, such as PDCD1 (PD-1), CD274 (PD-L1), CTLA4 and TIGIT. RIPK2 aggravated cytotoxic T lymphocyte (CTL) dysfunction and related to the poor efficacy of immune checkpoint blockade in skin cutaneous melanoma (SKCM) and kidney renal clear cell carcinoma (KIRC). High expression of RIPK2 promoted innate immunotherapy resistance and adaptive immunotherapy resistance through IL-6/JAK/STAT3 signaling, interferon-gamma response, and interferon-alpha response pathway.ConclusionsThese results confirmed that RIPK2 could serve as a prognostic biomarker and promoted immune therapy resistance via triggering cytotoxic T lymphocytes dysfunction.

MP06-13 TARGETING THE ANDROGEN RECEPTOR AXIS TO IMPROVE RESPONSE TO BLADDER CANCER IMMUNOTHERAPY

You have accessJournal of UrologyCME1 May 2022MP06-13 TARGETING THE ANDROGEN RECEPTOR AXIS TO IMPROVE RESPONSE TO BLADDER CANCER IMMUNOTHERAPY Typhaine Gris, Marjorie Besançon, France-Hélène Joncas, Valérie Picard, Alain Bergeron, Yves Fradet, and Paul Toren Typhaine GrisTyphaine Gris More articles by this author , Marjorie BesançonMarjorie Besançon More articles by this author , France-Hélène JoncasFrance-Hélène Joncas More articles by this author , Valérie PicardValérie Picard More articles by this author , Alain BergeronAlain Bergeron More articles by this author , Yves FradetYves Fradet More articles by this author , and Paul TorenPaul Toren More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002523.13AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Men are 3-4 times more likely to be diagnosed with bladder cancer (BCa) than women, who often have more aggressive tumors. Intravesical instillation of BCG is one of the first immunotherapies in widespread use and is given to patients with non-muscle invasive bladder cancer (NMIBC). Newer PD-1/PD-L1 checkpoint inhibitors now also have proven efficacy against BCa. Nonetheless, approximately 35% of patients fail to respond to BCG, with response rates to checkpoint immunotherapy much lower. Based on prior work suggesting the importance of the androgen receptor (AR) in BCa, we investigated AR-targeting as a novel strategy to improve the response to BCa immunotherapy in the MBT-2 murine BCa model. METHODS: Subcutaneous injection of 106 MBT-2 cells into male and female C3H mice was performed, with tumors harvested at tumor size limits. The AR antagonist enzalutamide (ENZ) was given in alone or combination with anti-PD-1 or intra-tumoral BCG treatments as previously described. Selected human NMIBC were collected following transurethral resection. After digestion of fresh human and murine tumors, flow cytometry analyses characterized the immune cells present. The anti-proliferative effect of AR antagonists against human and murine BCa cell lines was tested in vitro. RESULTS: The CD45+/CD45- ratio, extent of tumor infiltrating lymphocytes, and expression of immune checkpoint markers from the MBT-2 model is comparable to the immune composition of NMIBC tumors resected from patients. Moreover, the model also recapitulates previously observed human tumor immune cell gender differences. AR antagonist ENZ demonstrated anti-proliferative effects in lower grade human and murine BCa cell lines, while limited differences were seen in human high grade BCa cell lines. While ENZ alone had no effect in vivo, its use in combination with either anti-PD-1 or BCG treatment in male mice synergized to improve response rates. Complete response was confirmed by re-challenge of mice, demonstrating immune memory. Notably, the proportion of complete responses in male mice treated with the combination treatment is similar to that observed in female mice with either immunotherapy alone. Flow cytometry analyses showed that these therapeutic benefits in male mice are related to an increase in pro-inflammatory tumor infiltrating dendritic cells as well as a decrease in anti-inflammatory tumor infiltrating dendritic cells and myeloid-derived suppressor cells. CONCLUSIONS: Our results suggest that combining AR antagonism with immunotherapy in male BCa patients may potentiate the antitumor immune response and increase response rates. Accordingly, a Phase II trial to evaluate the efficacy of AR antagonism in combination with BCG to decrease NMIBC tumor recurrences in men is planned. Further, the MBT-2 murine model appears relevant to investigate immunologic BCa sex differences. Source of Funding: Cancer Research Society (Upcycle Grant) © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e82 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Typhaine Gris More articles by this author Marjorie Besançon More articles by this author France-Hélène Joncas More articles by this author Valérie Picard More articles by this author Alain Bergeron More articles by this author Yves Fradet More articles by this author Paul Toren More articles by this author Expand All Advertisement PDF DownloadLoading ...

Impact of Chemoembolic Regimen on Immune Cell Recruitment and Immune Checkpoint Marker Expression following Transcatheter Arterial Chemoembolization in a VX2 Rabbit Liver Tumor Model

PurposeTo characterize the effects of commonly used transcatheter arterial chemoembolization (TACE) regimens on the immune response and immune checkpoint marker expression using a VX2 rabbit liver tumor model. Materials and MethodsTwenty-four VX2 liver tumor-bearing New Zealand white rabbits were assigned to 7 groups (n = 3 per group) undergoing locoregional therapy as follows: (a) bicarbonate infusion without embolization, (b) conventional TACE (cTACE) using a water-in-oil emulsion containing doxorubicin mixed 1:2 with Lipiodol, drug-eluting embolic-TACE with either (c) idarubicin-eluting Oncozene microspheres (40 μm) or (d) doxorubicin-eluting Lumi beads (40–90 μm). For each therapy arm (b–d), a tandem set of 3 animals with additional bicarbonate infusion before TACE was added, to evaluate the effect of pH modification on the immune response. Three untreated rabbits served as controls. Tissue was harvested 24 hours after treatment, followed by digital immunohistochemistry quantification (counts/μm2 ± SEM) of tumor-infiltrating cluster of differentiation 3+ T-lymphocytes, human leukocyte antigen DR type antigen-presenting cells (APCs), cytotoxic T-lymphocyte–associated protein-4 (CTLA-4), and programmed cell death protein-1 (PD-1)/PD-1 ligand (PD-L1) pathway axis expression. ResultsLumi-bead TACE induced significantly more intratumoral T-cell and APC infiltration than cTACE and Oncozene-microsphere TACE. Additionally, tumors treated with Lumi-bead TACE expressed significantly higher intratumoral immune checkpoint markers compared with cTACE and Oncozene-microsphere TACE. Neoadjuvant bicarbonate demonstrated the most pronounced effect on cTACE and resulted in a significant increase in intratumoral cluster of differentiation 3+ T-cell infiltration compared with cTACE alone. ConclusionsThis preclinical study revealed significant differences in evoked tumor immunogenicity depending on the choice of chemoembolic regimen for TACE.

A risk model of 10 aging-related genes for predicting survival and immune response in triple-negative breast cancer.

Accumulated studies showed that the clinical significance of aging on the development and malignancy of tumors, while the relationship between aging and the prognosis, immune response in triple‐negative breast cancer (TNBC) has not been well clarified. Here, we constructed a risk model of 10 prognostic aging‐related genes (ARGs) from METABRIC database. Then, TNBC patients were classified into high‐ and low‐risk groups, the survival diversity, immune response, genomic function, and tumor mutation burden (TMB) between different risk groups were explored in METABRIC, TCGA, and GSE58812 cohorts. Results showed that patients in the high‐risk group had poorer survival outcomes compared to their counterparts (all p < 0.05), and the nomogram we established showed reliable prediction ability for survival in TNBC patients. Besides, TNBC patients with high‐risk scores had a lower expression of immune checkpoint markers and a lower fraction of activated immune cells. Furthermore, GSEA showed that Notch signaling pathway was significantly enriched in the high‐risk group. Thus, a risk model based on the aging‐related genes was developed and validated in this study, which may serve as a potential biomarker for prognosis and personalized treatment in TNBCs.

Pyroptosis-Related lncRNAs Predict the Prognosis and Immune Response in Patients With Breast Cancer.

Background: Breast cancer (BC) is the most common malignant tumor and the leading cause of cancer-related death in women worldwide. Pyroptosis and long noncoding RNAs (lncRNAs) have been demonstrated to play vital roles in the tumorigenesis and development of BC. However, the clinical significance of pyroptosis-related lncRNAs in BC remains unclear. Methods: Using the mRNA and lncRNA profiles of BC obtained from TCGA dataset, a risk model based on the pyroptosis-related lncRNAs for prognosis was constructed using univariate and multivariate Cox regression model, and least absolute shrinkage and selection operator. Patients were divided into high- and low-risk groups based on the risk model, and the prognosis value and immune response in different risk groups were analyzed. Furthermore, functional enrichment annotation, therapeutic signature, and tumor mutation burden were performed to evaluate the risk model we established. Moreover, the expression level and clinical significance of the selected pyroptosis-related lncRNAs were further validated in BC samples. Results: 3,364 pyroptosis-related lncRNAs were identified using Pearson’s correlation analysis. The risk model we constructed comprised 10 pyroptosis-related lncRNAs, which was identified as an independent predictor of overall survival (OS) in BC. The nomogram we constructed based on the clinicopathologic features and risk model yielded favorable performance for prognosis prediction in BC. In terms of immune response and mutation status, patients in the low-risk group had a higher expression of immune checkpoint markers and exhibited higher fractions of activated immune cells, while the high-risk group had a highly percentage of TMB. Further analyses in our cohort BC samples found that RP11-459E5.1 was significantly upregulated, while RP11-1070N10.3 and RP11-817J15.3 were downregulated and significantly associated with worse OS. Conclusion: The risk model based on the pyroptosis-related lncRNAs we established may be a promising tool for predicting the prognosis and personalized therapeutic response in BC patients.

Prognostic Autophagy-Related Model Revealed by Integrating Single-Cell RNA Sequencing Data and Bulk Gene Profiles in Gastric Cancer

Autophagy has been associated with tumor progression, prognosis, and treatment response. However, an autophagy-related model and their clinical significance have not yet been fully elucidated. In the present study, through the integrative analysis of bulk RNA sequencing and single-cell RNA sequencing, an autophagy-related risk model was identified. The model was capable of distinguishing the worse prognosis of patients with gastric cancer (GC), which was validated in TCGA and two independent Gene Expression Omnibus cohorts utilizing the survival analysis, and was also independent of other clinical covariates evaluated by multivariable Cox regression. The clinical value of this model was further assessed using a receiver operating characteristic (ROC) and nomogram analysis. Investigation of single-cell RNA sequencing uncovered that this model might act as an indicator of the dysfunctional characteristics of T cells in the high-risk group. Moreover, the high-risk group exhibited the lower expression of immune checkpoint markers (PDCD1 and CTLA4) than the low-risk group, which indicated the potential predictive power to the current immunotherapy response in patients with GC. In conclusion, this autophagy-associated risk model may be a useful tool for prognostic evaluation and will facilitate the potential application of this model as an indicator of the predictive immune checkpoint biomarkers.

The Effects of Differentially-Expressed Homeobox Family Genes on the Prognosis and HOXC6 on Immune Microenvironment Orchestration in Colorectal Cancer.

BackgroundThe homeobox (HOX) gene family encodes highly conserved transcription factors, that play important roles in the morphogenesis and embryonic development of vertebrates. Mammals have four similar HOX gene clusters, HOXA, HOXB, HOXC, and HOXD, which are located on chromosomes 7, 17,12 and 2 and consist of 38 genes. Some of these genes were found to be significantly related to a variety of tumors; however, it remains unknown whether abnormal expression of the HOX gene family affects prognosis and the tumor microenvironment (TME) reshaping in colorectal cancer (CRC). Therefore, we conducted this systematic exploration to provide additional information for the above questions.MethodsRNA sequencing data from The Cancer Genome Atlas (TCGA) and mRNA expression data from Gene Expression Omnibus (GEO) combined with online tumor analysis databases (UALCAN, TIMER, PrognoScan) were utilized to explore the relationship among abnormal expression of HOX family genes, prognosis and the tumor immune microenvironment in CRC.Results1. Differential expression and prognosis analysis: 24 genes were significantly differentially expressed in CRC compared to adjacent normal tissues, and seven upregulated genes were significantly associated with poor survival. Among these seven genes, univariate and multivariate Cox regression analysis revealed that only high expression of HOXC6 significantly contributed to poor prognosis; 2. The influence of overexpressed HOXC6 on the pathway and TME: High HOXC6 expression was significantly related to the cytokine pathway and expression of T cell attraction chemokines, the infiltration ratio of immune cells, expression of immune checkpoint markers, tumor mutation burden (TMB) scores and microsatellite instability-high (MSI-H) scores; 3. Stratified analysis based on stages: In stage IV, HOXC6 overexpression had no significant impact on TMB, MSI-H, infiltration ratio of immune cells and response prediction of immune checkpoint blockers (ICBs), which contributed to significantly poor overall survival (OS).ConclusionSeven differentially expressed HOX family genes had significantly worse prognoses. Among them, overexpressed HOXC6 contributed the most to poor OS. High expression of HOXC6 was significantly associated with high immunogenicity in nonmetastatic CRC. Further research on HOXC6 is therefore worthwhile to provide potential alternatives in CRC immunotherapy.

Different expression of immune checkpoint markers on bronchoalveolar lavage CD4+ cells: a comparison between hypersensitivity pneumonitis and sarcoidosis.

Different expression of immune checkpoint markers on bronchoalveolar lavage CD4+ cells: a comparison between hypersensitivity pneumonitis and sarcoidosis.